Research in Plant Disease (식물병연구)

The Korean Society of Plant Pathology (한국식물병리학회)
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Development of Rapid Immune-gold Strip Kit for On-Site Diagnosis of Tomato spotted wilt virus

  • Yoon, Ju-Yeon (Department of Horticulture, Biotechnology and Landscape, Seoul Women's University) ;
  • Choi, Gug-Seoun (Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Cho, In-Sook (Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, Rural Development Administration) ;
  • Choi, Seung-Kook (Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, Rural Development Administration)
  • Received : 2013.09.24
  • Accepted : 2014.03.04
  • Published : 2014.03.31
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Abstract

A rapid, user-friendly and simple immune-chromatographic dipstick kit named 'rapid immune-gold strip' (RIGS) kit was developed in a novel single strip format to detect on-site detection of Tomato spotted wilt virus (TSWV). Immunoglobulin G (IgG) from polyclonal antisera raised in rabbits against TSWV was purified through protein-A affinity chromatography and then the purified TSWV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TSWV antibody was used as a control line on the same strip. The diagnosis process with the TSWV-RIGS involves simply grinding the suspect plant sample in a bag that contains the extraction buffer and inserting the strip the bag. Results can be seen in 2-5 minutes. The flow of the complexes of gold particles coated with TSWV-IgG and a crude sap from TSWV-infected pepper, tobacco and tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TSWV. The RIGS-TSWV kit did not show any cross-reactions against other tomato-infecting viruses unrelated to TSWV. These results indicate that the TSWV-RIGS kit is highly sensitive and is not required for laboratory training and experience prior to testing. The TSWV-RIGS kit is suitable for on-site detection of suspect TSWV-infected plants as well as for laboratory diagnosis.

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References

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